Overcoming expression and purification problems of RhoGDI using a family of "parallel" expression vectors

Protein Expr Purif. 1999 Feb;15(1):34-9. doi: 10.1006/prep.1998.1003.


We describe the construction of expression vectors based on three of the most frequently used gene fusion affinity tags [glutathione S-transferase (GST), maltose binding protein (MBP), and the His6 peptide]. The polylinkers of pGEX4T1, pMal-c2, and a pET vector were replaced with the polylinker isolated from the baculovirus expression plasmid pFastBac. Once appropriate restriction sites have been introduced into a gene, it can be fused to all three affinity tags with little effort, allowing expression-screening experiments to be performed efficiently. We discuss the development and use of these vectors with respect to overcoming purification problems encountered for the RhoA GDP/GTP nucleotide dissociation inhibitor (RhoGDI) and their advantages over commercially available expression vectors.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • ATP-Binding Cassette Transporters*
  • Amino Acid Sequence
  • Baculoviridae
  • Base Sequence
  • Carrier Proteins
  • Chromatography, Affinity
  • Cloning, Molecular / methods
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli
  • Escherichia coli Proteins*
  • GTP-Binding Proteins / biosynthesis
  • GTP-Binding Proteins / genetics*
  • GTP-Binding Proteins / isolation & purification*
  • Genetic Vectors*
  • Glutathione Transferase
  • Guanine Nucleotide Dissociation Inhibitors*
  • Histidine
  • Humans
  • Maltose-Binding Proteins
  • Molecular Sequence Data
  • Monosaccharide Transport Proteins*
  • Mutagenesis, Site-Directed
  • Peptides
  • Point Mutation
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / isolation & purification
  • Restriction Mapping
  • rho Guanine Nucleotide Dissociation Inhibitor alpha
  • rho-Specific Guanine Nucleotide Dissociation Inhibitors


  • ARHGDIA protein, human
  • ATP-Binding Cassette Transporters
  • Carrier Proteins
  • Escherichia coli Proteins
  • Guanine Nucleotide Dissociation Inhibitors
  • Maltose-Binding Proteins
  • Monosaccharide Transport Proteins
  • Peptides
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • maltose transport system, E coli
  • rho Guanine Nucleotide Dissociation Inhibitor alpha
  • rho-Specific Guanine Nucleotide Dissociation Inhibitors
  • Histidine
  • Glutathione Transferase
  • GTP-Binding Proteins

Associated data

  • GENBANK/AF097413